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Abstract

In this paper we provide, for the first time to our knowledge, the effective orientation of the SHG source in cultured cortical neuronal processes in vitro. This is done by the use of the polarization sensitive second harmonic generation (PSHG) imaging microscopy technique. By performing a pixel-level resolution analysis we found that the SHG dipole source has a distribution of angles centered at θe =33.96°, with a bandwidth of Δθe = 12.85°. This orientation can be related with the molecular geometry of the tubulin heterodimmer contained in microtubules.

Fig. 4. (a) Resulting image of the b parameter for those pixels with r2>90% (scale bar is 10μm). (b)-(e) Histograms of the different parameters (within the same ROI of Fig. 4(a)) for b, E, δ and φ, respectively. The red line in each figure is a Gaussian fit to each of the histograms. (f) Examples of the fit of three different pixels. The retrieved parameters are: light grey line b=4.43, E=0.05, φ=2.64, δ=0; green dashed-line b=2.46, E=0.17, φ=7.44, δ=0; blue line b=7.77, E=0.02, φ=0, δ=0.09.

Tables (1)

Table 1. The values of the parameters θeδ, b, E, φ, of Eq. (3) The different values are the mean, and the standard deviation (2σ) of a Gaussian distribution fit for the obtained histograms with pixels with r2>90% in the ROI indicated in Fig. 4(a).

Metrics

Table 1.

The values of the parameters θeδ, b, E, φ, of Eq. (3) The different values are the mean, and the standard deviation (2σ) of a Gaussian distribution fit for the obtained histograms with pixels with r2>90% in the ROI indicated in Fig. 4(a).